Page 10 - S. Plant biology

[S. Plant biology-6]

Genome editing effect of the CRISPR/Cas9 system in hybrid

poplar using agrobacterium mediated transformation

Su Jin Park¹, Eun-Kyung Bae¹, Hyun-Ho Kim¹, Young-Im Choi¹˙*

¹Department of Forest Bioresources, National Institute of Forest Science, Suwon 16631, Korea

Engineered nucleases can effectively digest and mutate targeted genes in the plant genome. The CRISPR/CRISPR-
associated protein 9 (Cas9) system can induce gene editing at a high rate within months and can be effectively
used for long-lived trees. The first reported generation of an albino mutant tree using the CRISPR/Cas9 system

employed the agrobacterium-transformation method in Populus tomentosa. In addition, PDS gene mutation by

CRISPR/Cas9 has been achieved in various fruit trees. We attempted to determine whether the CRISPR/Cas9 system
can induce PDS gene mutation in the hybrid poplar (Populus alba × P. glandulosa). The phenotype and DNA
sequence mutations were analyzed in plants with mutated PagPDS1 genes induced by pHSE/Cas9-PagPDS1 gRNA

from the transgenic hybrid poplar after agrobacterium-mediated transformation. The mutation rates of the albino,
pale green, and green phenotypes were 60%, 14.55%, and 25.45%, respectively. Collectively, the results show that

the CRISPR/Cas9 system was effective in reducing gene expression and altering the plant phenotype by editing a
hybrid poplar target gene with two alleles. We expect that this technique will be used to improve tree quality.